Solid matrix for the storage of biological samples

ABSTRACT

The present invention relates to a method for storage and subsequent lysis of a sample in which the sample is immobilized on a solid support. The solid matrix is embedded with a low concentration of both a chaotropic salt and a surfactant which act synergistically to efficiently store and lyse a biological sample.

FIELD OF INVENTION

The present invention relates to the field of sample collection fornucleic acid amplification, particularly to the use of the polymerasechain reaction to amplify nucleic acids. The invention provides methodsand kits which can be used to collect cell samples containing nucleicacids by reducing the use of chemicals that may interfere with thepolymerase chain reaction. The invention has applications in thelong-term storage, recovery and further processing of nucleic acids andis particularly useful in genotyping, diagnostics and forensicsapplications.

BACKGROUND OF THE INVENTION

Long-term storage, transport and archiving of nucleic acids on filterpaper or chemically modified matrices is a well-known technique forpreserving genetic material before the DNA or RNA is extracted andisolated in a form for use in genetic analysis such as PCR. Thus, EP1563091 (Smith et al, Whatman) relates to methods for storing nucleicacids from samples such as cells or cell lysates. The nucleic acid isisolated and stored for extended periods of time, at room temperatureand humidity, on a wide variety of filters and other types of solidsupport or solid phase media. Moreover, the document describes methodsfor storing nucleic acid-containing samples on a wide range of solidsupport matrices in tubes, columns, or multiwell plates.

WO 90/03959 (Burgoyne) describes a cellulose-based solid support for thestorage of DNA, including blood DNA, comprising a solid matrix having acompound or composition which protects against degradation of DNAincorporated into or absorbed on the matrix. This document alsodiscloses methods for storage of DNA using the solid medium, and forrecovery of or in situ use of DNA.

U.S. Pat. No. 5,705,345 (Lundin et al.) describes a method of nucleicacid preparation whereby the sample containing cells is lysed to releasenucleic acid and the sample is treated with cyclodextrin to neutralizethe extractant. The advantage of this system is that conventionaldetergent removal requires a separation step however with the additionof cyclodextrin to neutralize the detergent it would remove theseparation step needed and reduce chance of contamination.

U.S. Pat. No. 5,496,562 (Burgoyne) describes a cellulose-based solidmedium and method for DNA storage. Method for storage and transport ofDNA on the solid medium, as well as methods which involve either (a) therecovery of the DNA from the solid medium or (b) the use of the DNA insitu on the solid medium (for example, DNA sequence amplification byPCR) are disclosed. The method described incorporates a surfactant ordetergent on the surface of the solid medium and would require aseparate step for the removal of the detergent before PCR is performedas a detergent would interfere with the PCR process.

EP 2290099 B1 (Qiagen) describes again a method for processing andamplifying DNA. The method includes the steps of contacting the samplecontaining DNA to a solid support wherein a lysis reagent is bound tothe solid support. The DNA is subsequently treated with a DNA purifyingreagent and is purified.

WO96/39813 (Burgoyne) describes a solid medium for storing a sample ofgenetic material and subsequent analysis; the solid medium comprising aprotein denaturing agent and a chelating agent. The method described isfor chelating agents which are any compound capable of complexingmultivalent ions including Group II and Group III multivalent metal ionsand transition metal ions.

WO 96/18731 (Deggerdal) describes a method of isolating nucleic acidwhereby the sample is bound to a solid support and the sample iscontacted with a detergent and subsequent steps performed to isolate thenucleic acid.

WO 00/53807 (Smith, Whatman) describes a medium for the storage andlysis of samples containing genetic material which can be eluted andanalyzed. The medium is coated with a lysis reagent. In addition themedium could be coated with a weak base, a chelating agent, a surfactantand optionally uric acid.

WO 99/38962 (Health, Gentra Systems Inc.) describes a solid support witha bound lysis reagent. The lysis reagent can comprise of a detergent, achelating agent, water and optionally an RNA digesting enzyme. The solidsupport again would require further steps for purification of thenucleic acid for amplification analysis.

Inhibitors of PCR

There are significant problems with inhibitory substances or inhibitorsinterfering with polymerase chain reactions on nucleic acids, includingnucleic acids stored or immobilized on solid supports such ascellulose-derived filters. Sources of inhibitors can be the materialsand reagents such as detergents that come into contact with samplesduring processing or nucleic acid purification. These include excessKCl, NaCl and other salts, ionic detergents such as sodium deoxycholate,sarkosyl and sodium dodecyl sulphate (SDS), ethanol and isopropanol,phenol (see Burgoyne WO 90/03959) and others. All users of PCR arelikely to be impacted by inhibitors at some time, but the wide range offorensic sample types and variety of sampling conditions encounteredmake forensic scientists particularly vulnerable.

The presence of chemicals that interfere with PCR analysis in samples iswell-known and a number of approaches are documented that describe theirremoval. However, none of these approaches are particularly efficient.In particular the application of samples to solid supports, particularlycellulose-derived materials, increases the problems with PCR inhibitionbecause of the inclusion of potent chemicals such as SDS and chaotropicsalts which are typically used to stabilize nucleic acids on the solidsupport (see Burgoyne WO 90/03959).

PCR inhibitors usually affect PCR through interaction with DNA orinterference with the DNA polymerase. In a multiplex PCR reaction, it ispossible for the different sequences to experience different inhibitioneffects to varying degrees, leading to a disparity in results.Alternatively, various inhibitors reduce the availability of cofactors(such as magnesium) or otherwise interfere with the interaction of PCRcofactors with the DNA polymerase. Some of the pitfalls of quantitativereal-time reverse transcription polymerase chain reaction, including theeffect of inhibitors, are described by Bustin & Nolan (J. BiomolecularTechniques, 2004, 15, 155-166).

Current methods for inhibitor removal are often carried out during theDNA purification procedure by binding to single or double stranded DNAcovalently linked to a support. However, this is a tedious process andprior art methods have a number of clear disadvantages in terms of cost,complexity and in particular, user time.

U.S. Pat. No. 5,705,345 (Lundin et al.) describes a method of nucleicacid preparation whereby the sample containing cells is lysed to releasenucleic acid and the sample is treated with cyclodextrin to neutralizethe extractant. The advantage of this system is that conventionaldetergent removal requires a separation step however with the additionof sequestrants such as cyclodextrin to neutralize the detergent itwould remove the separation step needed and reduce chance ofcontamination.

WO 2010/066908 (Beckers et al.,) describes the use of cyclodextrins toimprove the specificity, sensitivity and/or yield of PCR. The methodclaimed is an amplification reaction which is performed in a reactionmixture comprising at least one cyclodextrin and performing theamplification reaction on said reaction.

Alternative methods involve the binding of nucleic acids in the presenceof chaotropic agents such that DNA binds to silica or glass particles orglass beads. This property was used to purify nucleic acid using glasspowder or silica beads under alkaline conditions. Typical chaotropicagents include guanidinium thiocyanate or guanidinium hydrochloride andrecently glass beads have been substituted with glass containingminicolumns. While the chaotropic agent may neutralize the interferenceof the detergent on the PCR process, the chaotropic agent itself mayreduce the efficiency of PCR amplification.

Wang et al. (Enzyme Microbiology Technology, 1990) describes a method ofrecovering protein from a bacteria by exposing E. coli to certaincombinations of guanidine and Triton X-100 in a solution. It was notedthat neither of the chemicals alone and at low concentrations extractsmore than 10% of intracellular protein however together were able torelease over 50% overall protein.

U.S. Pat. No. 5,939,259 (Schleicher & Schuell Inc.) describes a methodfor the collection and storage of nucleic acids by coating an absorbentmaterial with a solution containing from 0.5M to 2.0M chaotropic saltand allowed to dry. Other ingredients can be added to the invention inorder to enhance lysis or disruption of intact cells, bacteria andviruses which include anionic, cationic or zwitterionic surfactants,such as Tween 20 or Triton X-100.

Given the wide range of PCR inhibitor-laden sample types and the optionsavailable for handling them, a multifaceted approach is the bestsolution for amplification failure. There is a need to reduce anypotential PCR inhibitors used in the nucleic acid collection processwherein the nucleic acid is immobilized on a solid support and therebyimprove PCR amplification.

The present invention addresses this problem and provides methods andkits which can be used for the collection of nucleic acids using solidsupports, particularly cellulose-derived supports.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows the conductivity versus concentration of the guanidinethiocyanate treated ETF paper.

FIG. 2 shows the conductivity versus concentration of the guanidinethiocyanate and Triton X100 treated ETF paper.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides a solid matrix which can be used to storeand lyse biological samples by contacting a solid support embedded witha surfactant and a chaotropic salt.

According to a first aspect of the present invention, there is provideda solid matrix for storing and/or lysing a biological sample comprisinga surfactant and a chaotropic salt, wherein the concentration of thechaotropic salt is less than 0.5M.

The advantage of embedding a surfactant and a chaotropic salt onto asolid matrix for sample collection is the two chemicals can reactsynergistically to lyse the cell thereby allowing the use of a lowerconcentration of both chemicals. The benefits of using a lowerconcentration of surfactant and a chaotropic salt on a solid matrix isto reduce interference of these two chemicals in the PCR process.

In one aspect, the solid matrix wherein the chaotropic saltconcentration is greater than 0.05M.

In another aspect, the solid matrix wherein the surfactant concentrationis greater than 0.05M.

In a further aspect, the solid matrix wherein the surfactantconcentration is less than 0.5M.

In a further aspect, the solid matrix wherein the chaotropic salt andsurfactant is present in a ratio of 1:1.

In another aspect the solid matrix wherein the chaotropic salt andsurfactant is present in a ratio of 1:10.

In another aspect, the solid matrix wherein the chaotropic salt andsurfactant is present in a ratio of 10:1.

In a further aspect, the solid matrix wherein the concentration ofchaotropic salt to surfactant is selected from the group consisting of0.01M:01M, 0.1M:0.01M, 0.05M:0.05M, 0.1M:0.1M, 0.2M:0.2M, 0.3M:0.3M and0.4M:0.4M.

In a further aspect the solid matrix wherein said surfactant is TritonX100.

In a further aspect the solid matrix wherein the chaotropic salt is aguanidinium salt.

In one aspect, the solid matrix wherein the chaotropic salt is selectedfrom the group consisting of guanidine hydrochloride, guanidineisothiocyanate and guanidine thiocyanate.

In a further aspect, the solid matrix wherein the chaotropic salt isguanidine thiocyanate.

In another aspect, the solid matrix wherein the biological sample isselected from the group consisting of a cellular sample, blood, serum,semen, cerebral spinal fluid, synovial fluid, lymphatic fluid, saliva,buccal, cervical cell, vaginal cell, urine, faeces, hair, skin andmuscle.

In another aspect, the solid matrix wherein the solid matrix is selectedfrom the group consisting of: alginate, glass, glass fiber, glassmicrofiber, silica, silica gel, silica oxide, cellulose, nitrocellulose,carboxymethylcellulose, polyester, polyamide, carbohydrate polymers,polypropylene, polytetrafluororethylene, polyvinylidinefluoride, wooland porous ceramics.

In another aspect, the solid matrix wherein the solid matrix is acellulose based matrix.

In another aspect, the solid matrix wherein said cellulose based matrixis in the form of a pre punched disc.

In a further aspect the solid matrix additionally wherein said cellulosebased matrix is in the form of an FTA pre punched disc.

The solid matrix may comprise a glass or silica-based solid phasemedium, a plastics-based solid phase medium or a cellulose-based solidphase medium. The solid support is preferably a cellulose-based matrix.Examples of cellulose-based matrices include FTA™ (data file 51668), 903neonatal cards and 31-ETF cards available from GE Healthcare.

In a further aspect, said solid matrix is impregnated with sodiumdodecyl sulfate (SDS), ethylenediaminetetracetic acid (EDTA) and uricacid.

According to a second aspect of the present invention, there is provideda method of storing and/or lysing a biological sample comprising thesteps;

i) contacting a solid matrix of any preceding claim with a biologicalsample,

ii) storing said solid matrix with said biological sample,

wherein the chaotropic salt and surfactant have a synergistic effect inlysing the cell.

In a further aspect, the method wherein the method comprises a furtherstep iii) of recovering the cell lysate from the solid matrix.

In another aspect, the method wherein the biological sample is selectedfrom a group comprising consisting of a cellular sample, blood, serum,semen, cerebral spinal fluid, synovial fluid, lymphatic fluid, saliva,buccal, cervical cell, vaginal cell, urine, faeces, hair, skin andmuscle. The biological sample may originate from a mammal, bird, fish orplant or a cell culture thereof. Preferably the biological sample ismammalian in origin, most preferably human in origin. The samplecontaining the nucleic acid may be derived from any source. Thisincludes, for example, physiological/pathological body fluids (e.g.secretions, excretions, exudates) or cell suspensions of humans andanimals; physiological/pathological liquids or cell suspensions ofplants; liquid products, extracts or suspensions of bacteria, fungi,plasmids, viruses, prions, etc.; liquid extracts or homogenates of humanor animal body tissues (e.g., bone, liver, kidney, etc.); media from DNAor RNA synthesis, mixtures of chemically or biochemically synthesizedDNA or RNA; and any other source in which DNA or RNA is or can be in aliquid medium.

In one aspect, the biological sample is immobilised on the solid supportfor at least 24 hours. The biological sample may be immobilised on thesolid support for longer periods, for example, for at least 7 days, forat least 30 days, for at least 90 days, for at least 180 days, for atleast one year, and for at least 10 years. In this way the biologicalsample may be stored in a dried form which is suitable for subsequentanalysis. Typically, samples are stored at temperatures from −200° C. to40° C. In addition, stored samples may be optionally stored in dry ordesiccated conditions or under inert atmospheres.

According to a third aspect of the present invention, there is provideda method of manufacturing a solid matrix comprising the steps,

-   i) dipping a solid matrix into a mixture of a surfactant and a    chaotropic salt wherein the concentration of said chaotropic salt is    less than 0.5M, and-   ii) allowing the solid matrix to dry.

In one aspect, the method wherein the concentration of said chaotropicsalt is greater than 0.05M

In another aspect, the method wherein said surfactant concentration isgreater than 0.05M and is less than 0.5M.

In a further aspect, the method wherein said surfactant concentration isgreater than 0.05M and is less than 0.5M.

In another aspect, the method wherein the chaotropic salt and surfactantis present in a ratio selected from the group consisting of 1:1, 1:10,10:1.

In another aspect, the method wherein the concentration of chaotropicsalt to surfactant is selected from the group consisting of 0.01M:01M,0.1M:0.01M, 0.05M:0.05M, 0.1M:0.1M, 0.2M:0.2M, 0.3M:0.3M and 0.4M:0.4M.

In a further aspect, the method wherein said surfactant is Triton X100.

In another aspect, the method wherein the chaotropic salt is selectedfrom the group consisting of guanidine hydrochloride, guanidineisothiocyanate and guanidine thiocyanate.

In another aspect, the method wherein the solid matrix is selected fromthe group consisting of: alginate, glass, glass fiber, glass microfiber,silica, silica gel, silica oxide, cellulose, nitrocellulose,carboxymethylcellulose, polyester, polyamide, carbohydrate polymers,polypropylene, polytetrafluororethylene, polyvinylidinefluoride, wool orporous ceramics.

According to a fourth aspect, a kit for storing and amplifying nucleicacid comprising a solid matrix as described herein and instructions foruse thereof.

Chemicals and Materials Used

A list of the chemicals and their sources is given below:

31-ETF paper was obtained from GE Healthcare UK Limited;

Guanidinium thiocyanate (Sigma-Aldrich G9277)

Triton X100 (Sigma-Aldrich T8787)

Sterile water (Sigma W4502).

Coating tray

Glass Dipping tray

Mueller-Hinton-agar plates (“ready to use” plates of mibius—P05007Acontain the agar of Oxoid—CM0337)

Sterile Rotilabo—swabs (EH 12.1 of Roth)

Sterile petri plates 80 mm

7 mm Harris Uni-core punch, 1.2 mm (Sigma, Catalogue numberZ708860-25ea, lot 3110);

Forceps

Positive 31-ETF control from previous lots of FTA®-treated paper

TSB (Oxoid Tryptone Soy Broth, Art-Nr. CM0129)—150 ml in Erlenmeer flaskfor culture suspension.

95% and 70% Ethyl alcohol

Staphylococcus aureus (ATCC 25923 strain)

Measuring ruler

Experimental Results

ETF paper was coated in a mixture of Guanidine Thiocyanate and TritonX100 using a hand dipping technique. Several mixtures of various molarconcentrations of Guanidine Thiocyanate: Triton X100 was prepared.

An example of the preparation method used to produce the mixture ofGuanidine Thiocyanate: Triton X100 was as follows:

For a 0.5M:0.5M Guanidine Thiocyanate: Triton X100 solution 32.5 g ofTriton X100 was slowly added to 100 mL of dH₂O, using a dropping funnel,due to the low solubility of Triton X100 in water the solutions wascontinuously stirred as the reagent was added to the water. Once theTriton X100 had dissolved 5.908 g of Guanidine Thiocyanate was slowlyadded and the solution was stirred to ensure a homogenous solution. TheETF paper was dipped into the homogenous solution and allowed to dry.Table 1 below shows the concentrations and weight of GuanidineThiocyanate and Triton X100 used in the different sample preparations.

TABLE 1 Weight of Guanidine Thiocyanate:Triton X100 Weight of TritonX100 Thiocyanate used in molar concentrations used in grams grams 0.5M32.5 5.908 0.4M 26.0 4.726 0.3M 19.5 3.544 0.2M 13.0 2.363 0.1M 6.51.181 0.01M  0.65 0.118

The coated ETF paper samples were measured for the zone of inhibition(ZOI). A visible zone of inhibition, described as the area encompassingthe punch where bacterial growth is inhibited, is used to assess theanti-microbial action of the FTA treated paper.

A day before the ZOI test the Staphylococcus aureus suspension wasprepared in a 150 mL simple concentrates Tryptone Soy Broth. The culturesuspension flask was incubated at 37° C. after the inoculation 18±2.Three punches were taken from each individual ETF treated sample. The 7mm punch was rinsed with 95% EtOH after each punch was taken. Thepunches were placed in a sterile plate. The negative control was anuntreated ETF paper. A sample from a previous lot of ETF treated paperthat produced a well-defined ZOI was used as the positive control.

A germ count was performed for a growth confirmation of the culturesuspension. The culture suspension had a minimum of 10⁸ CFU/mL for thetest to count. The culture suspension was applied on to a single agarplate with a sterile swab. For each tested sample a minimum of 3 plateswere prepared. Using forceps and aseptic technique, the paper puncheswere placed onto the appropriate Mueller-Hinton agar plate—each platecontained a positive and a negative sample, along with 3 punches of thesame sample paper. The plates were incubated in an aerobic incubator at37° C. and were examined after 16 to 18 hours of incubation and a secondtime when necessary. The ZOI was calculated using a ruler to measure thediameter (including the 7 mm disc) of each zone of complete inhibitionto the nearest whole millimeter. The area showing no macroscopicallyvisible growth is defined as the end point.

The results showing the ZOI of ETF coated with varying concentrations ofGuanidine Thiocyanate and Triton X100 are shown in table 2 and table 3below.

TABLE 2 Molar Concentration of Guanidine:Triton Bacteria free X100areola in mm 0.5M:0.5M 11 11 11 0.4M:0.4M 10 11 11 0.3M:0.3M 11 11 110.2M:0.2M 11 11 11 0.1M:0.1M 11 11 11 0.01M:0.01   0 0 0

TABLE 3 Guanidine Triton X100 Bacteria free Number conc M conc M areolain mm 1 0.01 0.1 0 0 0 2 0.1 0.01 8 8 8 3 0.01 0.1 0 0 0 4 0.01 0.01 0 00 5 0.055 0.055 8 8 8 6 0.1 0.1 8 8 8 7 0.1 0.01 8 8 8 8 0.055 0.055 8 88 9 0.1 0.1 8 8 10 10 0.01 0.01 0 0 0 11 0.01 0.01 0 0 0 12 0.1 0.01 8 88 13 0.01 0.1 8 8 8 14 0.1 0.1 8 8 8 15 0.055 0.055 8 8 8

As a control individual components of the cell lysing agents Triton X100and Guanidine Thiocyanate were separately used to coat ETF and the ZOIwas tested as described above. The results showing the ZOI of ETFtreated with Guanidine Thiocyanate or Triton X100 alone are shown intable 4 below.

TABLE 4 Molar Concentration Bacteria free of cell lysing agent areola inmm 0.1M Guanidine 0 0 0 0.1M Triton X100 0 0 0

The concentration of Guanidine Thiocyanate and Triton X100 that wastaken up by the ETF paper was calculated using conductivity analysis. Acalibration curve of concentration versus conductivity yielded an R² of0.9945. Using the data from the conductivity experiment theconcentration of guanidine present on the hybrid substrates wasinferred.

A 7 mm punch of ETF paper was placed in a test tube and 20 mL of H2O wasadded. The solution was shaken for 5 minutes and left to stand, theconductivity meter was calibrated and the probe placed in the test tube,slightly agitated and the reading taken. The conductivity over the rangeof concentrations coated onto the ETF paper was analysed and the resultsare shown in Table 5 below and the results were plotted on a graph shownin FIG. 1.

TABLE 5 Coating concentration Moles (guanidine thiocyanate on hybridpaper) Conductivity mS 0.1M 0.928 0.2M 1.541 0.3M 1.917 0.4M 2.573 0.5M2.802

The conductivity of ETF coated with Guanidine Thiocyanate and TritonX100 was determined and shown in FIG. 2. Using the establishedconductivity curve from the previous study an estimated coatingconcentration for guanidine on ETF was inferred and shown in table 6below.

TABLE 6 Coating concentration Actual concentration in Moles (guanidineon derived calculate using hybrid paper) plot formula % W/V 0.1M = 1.18%W/V 0.030 0.2M = 2.36% W/V 0.112 0.3M = 3.54% W/V 0.161 0.4M = 4.72% W/V0.272 0.5M = 5.90% W/V 0.279

Table 7 below represents the extracted data obtained from theconductivity analysis, the Triton X100 weight was calculated bysubtracting the Theoretical Guanidine Thiocyanate weight on the ETFpaper from the actual total weight increase of the Guanidine Thiocyanateand Triton 100X coated paper.

TABLE 7 Coating concen- Actual Theoretical Weight tration concentrationGuanidine increase in Triton in Moles derived weight grams of the X100(guanidine calculate using present Guanidine:Triton weight on hybridplot formula on each X100 present on paper) % W/V paper (g) mixturepaper (g) 0.1 0.030 0.020 0.1431 0.1231 0.2 0.112 0.027 0.1987 0.17170.3 0.161 0.045 0.3231 0.2781 0.4 0.272 0.061 0.4421 0.3811 0.5 0.2790.066 0.4715 0.4055

While preferred illustrative embodiments of the present invention aredescribed, one skilled in the art will appreciate that the presentinvention can be practiced by other than the described embodiments,which are presented for the purposes of illustration only and not by wayof limitation. The present invention is limited only by the claims thatfollow.

What is claimed is:
 1. A solid matrix for storing and/or lysing abiological sample comprising a surfactant and a chaotropic salt, whereinthe concentration of the chaotropic salt to surfactant is 0.1M:0.1M, thesurfactant is Triton X100, and the chaotropic salt is a guanidiniumsalt.
 2. The solid matrix of claim 1, wherein the biological sample isselected from the group consisting of a cellular sample, blood, serum,semen, cerebral spinal fluid, synovial fluid, lymphatic fluid, saliva,buccal, cervical cell, vaginal cell, urine, faeces, hair, skin andmuscle.
 3. The solid matrix of claim 1, wherein the solid matrix isselected from the group consisting of alginate, glass, glass fiber,glass microfiber, silica, silica gel, silica oxide, cellulose,nitrocellulose, carboxymethylcellulose, polyester, polyamide,carbohydrate polymers, polypropylene, polytetraflurorethylene,polyvinylidinefluoride, wool and porous ceramics.
 4. The solid matrix ofclaim 1, wherein the solid matrix is a cellulose based matrix.
 5. Thesolid matrix of claim 4, additionally wherein said cellulose basedmatrix is in the form of a pre punched disc.
 6. The solid matrix ofclaim 4, additionally wherein the cellulose based matrix is in the formof a pre punched disc.
 7. A method of storing and/or lysing a biologicalsample comprising the steps: i) contacting a solid matrix of anypreceding claim with a biological sample; and ii) storing said solidmatrix with said biological sample, wherein the concentration of thechaotropic salt to surfactant is 0.1M:0.1M, the surfactant is TritonX100, and the chaotropic salt is a guanidinium salt.
 8. The method ofclaim 7, further comprising a step iii) of recovering the cell lysatefrom the solid matrix.
 9. The method of claim 7, wherein the biologicalsample is selected from a group consisting of a cellular sample, blood,serum, semen, cerebral spinal fluid, synovial fluid, lymphatic fluid,saliva, buccal, cervical cell, vaginal cell, urine, faeces, hair, skinand muscle.
 10. A method of manufacturing a solid matrix comprising thesteps: i) dipping a solid matrix into a mixture of a surfactant and achaotropic salt wherein the concentration of said chaotropic salt tosurfactant is 0.1M:0.1M, the surfactant is Triton X100, and thechaotropic salt is a guanidinium salt; and ii) allowing the solid matrixto dry.
 11. The method of claim 10, wherein the solid matrix is selectedfrom the group consisting of alginate, glass, glass fiber, glassmicrofiber, silica, silica gel, silica oxide, cellulose, nitrocellulose,carboxymethylcellulose, polyester, polyamide, carbohydrate polymers,polypropylene, polytetraflurorethylene, polyvinylidinefluoride, wool orporous ceramics.
 12. A kit for storing and amplifying nucleic acidcomprising the solid matrix of claim 1 and instructions for use thereof.